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SUMOylation of Pdia3 exacerbates proinsulin misfolding and ER stress in pancreatic beta cells

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Abstract

SUMOylation has long been recognized to regulate multiple biological processes in pancreatic beta cells, but its impact on proinsulin disulfide maturation and endoplasmic reticulum (ER) stress remains elusive. Herein, we conducted comparative proteomic analyses of SUMOylated proteins in primary mouse/human islets following proinflammatory cytokine stimulation. Cytokine challenge rendered beta cells to undergo a SUMOylation turnover manifested by the changes of SUMOylation substrates and SUMOylation levels for multiple substrates. Our data support that SUMOylation may play a crucial role to regulate proinsulin misfolding and ER stress at least by targeting Protein Disulfide Isomerase a3 (Pdia3). SUMOylation regulates Pdia3 enzymatic activity, subcellular localization, and protein binding ability. Furthermore, SUMOylation of Pdia3 exacerbated proinsulin misfolding and ER stress, and repressed Stat3 activation. In contrast, disruption of Pdia3 SUMOylation markedly rescued the outcomes. Collectively, our study expands the understanding how SUMOylation regulates ER stress in beta cells, which shed light on developing potential strategies against beta cell dysfunction.

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Data availability

All data needed to evaluate the conclusion in the paper are present in the paper and/or the electronic supplementary material. Additional data related to this paper may be requested from the authors.

Abbreviations

C:

cysteine

Co-IP:

co-immunoprecipitation

CRP:

C-reactive protein

DTT:

dithiothreitol

ECAR:

extracellular acidification rate

GSIS:

glucose-stimulated insulin secretion

K:

lysine

MU:

mutant

OCR:

oxygen consumption rate

PDI:

protein disulfide isomerase

PTM:

post-translational modification

R:

arginine

RLU:

relative luciferase

SUMO:

small ubiquitin like modifier

Thap:

thapsigargin

UBC9:

SUMO-conjugating enzyme 2

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Acknowledgments

We sincerely thank Dr. Jinxiu Li (Shenzhen Third People’s Hospital, Shenzhen, Guangdong, China) for discussing and reviewing the manuscript.

Code availability

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Funding

This study was supported by the Ministry of Science and Technology of China (2016YFC1305002 and 2017YFC1309603), the National Natural Science Foundation of China (81471046, 81530024, 91749207, 81920108009, 81770823 and 81670729), NHC Drug Discovery Program of China (2017ZX09304022–07), the Department of Science and Technology of Hubei Province (2017ACA096), the Integrated Innovative Team for Major Human Disease Programs of Tongji Medical College, Huazhong University of Science and Technology, and the Innovative Funding for Translational Research from Tongji Hospital.

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Authors and Affiliations

  1. The Center for Biomedical Research, Tongji Hospital Research Building, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430000, China

    Na Li, Xi Luo, Qilin Yu, Ping Yang, Zhishui Chen, Xinqiang Wang, Jipin Jiang, Jing Xu, Fei Xiong, Shu Zhang & Cong-Yi Wang

  2. Key Laboratory of Organ Transplantation, Ministry of Education, NHC Key Laboratory of Organ Transplantation, Key Laboratory of Organ Transplantation, Chinese Academy of Medical Sciences, Tongji Hospital Research Building, Tongji Hospital, Wuhan, China

    Zhishui Chen, Xinqiang Wang, Jipin Jiang, Jing Xu, Fei Xiong, Shu Zhang & Cong-Yi Wang

  3. Clinical Molecular Immunology Center, Department of Immunology, School of Medicine, Yangtze University, Jingzhou, 434023, China

    Quan Gong

  4. ULB Center for Diabetes Research, Université Libre de Bruxelles, 808 Route de Lennik, B-1070, Brussels, Belgium

    Decio L. Eizirik

  5. Indiana Biosciences Research Institute (IBRI), Indianapolis, IN, USA

    Decio L. Eizirik

  6. Diabetes Center, the Second Xiangya Hospital, Institute of Metabolism and Endocrinology, Central South University, Changsha, China

    Zhiguang Zhou

  7. Department of Endocrinology and Metabolism, Shandong Provincial Hospital affiliated to Shandong First Medical University, Jinan, China

    Jiajun Zhao

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  1. Na Li
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Contributions

N.L. and X.L. were responsible for conducting all experiments, data analysis, and writing the manuscript. F.X., Q.Y., and P.Y. were responsible for conducting all experiments and data analysis. N.L., S.Z., and C.-Y.W. were involved in all method establishment and modification. X.W., J.J., and J.X. were responsible for collecting human pancreas. N.L. and C.-Y.W were responsible for writing the manuscript. S.Z. and Q.G. contributed to discuss and review the manuscript. N.L. and C.-Y.W. were responsible for funding acquisition. D.L E., Z.Z., J.Z., and S.Z. contributed to the review of the manuscript and manuscript preparation. C.-Y.W. is the guarantor of this work and had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.

Corresponding authors

Correspondence to Fei Xiong, Shu Zhang or Cong-Yi Wang.

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Li, N., Luo, X., Yu, Q. et al. SUMOylation of Pdia3 exacerbates proinsulin misfolding and ER stress in pancreatic beta cells. J Mol Med 98, 1795–1807 (2020). https://doi.org/10.1007/s00109-020-02006-6

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